Objective：To construct recombinant PNPLA7 shRNA adenovirus and check its knockdown efficiency in liver. Provide a new approach for further investigating the role of PNPLA7 on hepatic triglyceride metabolism. Methods： siNC and siPNPLA7 were designed and cloned into the p-shuttle-CMV-silence vector. After Sequencing identification，the plasmid was linearized by PmeⅠ and recombined with backbone pAdeasy in BJ5183 competent cells. The recombinant plasmid was linearized by PacⅠand transfected into 293A cells for packaging Ad-shPNPLA7 and Ad-shNC adenovirus. After 7 days of tail vein injection，Western blot was applied to determine the knockdown efficiency of pAdeasy-shPNPLA7 adenovirus in liver，and hepatic triglycerides content were measured. Results：The hepatic PNPLA7 protein levels were significantly decreased while the hepatic TAG levels were significantly increased in mice with tail vein injection of pAdeasy-shPNPLA7 adenovirus compared with control mice. Conclusion：The recombinant adenovirus of pAdeasy-shPNPLA7 and pAdeasy-shNC were successfully constructed. The PNPLA7 may play a role in hepatic triglycerides metabolism.